Chetomin

Cells were treated with different concentrations of anti-APO-1 antibodies,^{61 (link)} SuperKiller-TRAIL (Enzo Life Sciences, New York, NY, USA), etoposide (Sigma-Aldrich, St Louis, MO, USA), doxorubicin (Sigma-Aldrich) or chetomin (Enzo Life Sciences) for different time periods as indicated in the figure legends. Apoptotic cell death was examined by the analysis of DNA fragmentation according to the method of Nicoletti as described previously.^{8 (link)} Results are presented as the percent of specific DNA fragmentation using the formula: (percentage of experimental apoptosis−percentage of spontaneous apoptosis)/(100−percentage of spontaneous apoptosis) × 100.

Cilostazol (OPC-13013) [6-[4-(1-cyclohexyl-1H-tetrazol-5-yl)butoxy]-3,4-dihydro-2-(1H)-quinolinone] was donated by Otsuka Pharmaceutical Co. Ltd. (Tokushima, Japan), and was dissolved in dimethyl sulfoxide to produce a 10 mM stock solution. Recombinant Human HMGB1 was obtained from R&D Systems (Minneapolis, MN), and chetomin from Enzo Life Sciences (Plymouth Meeting, PA; purity >98% by HPLC analysis). BAY11-7082 [(E)3-[(4-methylphenyl)-sulfonyl]-2-propenenitrile], KT5720, resveratrol, and glycyrrhizin (a glycoconjugated triterpene produced by Glycyrrhiza glabra (licorice) from Sigma (St. Louis, MO). YC-1 [3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole] was from Cayman Chemical (Ann Arbor, MI] and dissolved in 1% DMSO. CD31 polyclonal antibody (1∶2,000 dilution; Abcam, Cambridge, UK) and monoclonal anti-vWF antibodies (1∶300 dilution; Dako, Glostrup, Denmark) were used to evaluate blood vessel numbers.